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在旧金山湾-三角洲地区,用于检测群落水平鱼类捕食的 10 种实时 PCR 检测方法。

Ten real-time PCR assays for detection of fish predation at the community level in the San Francisco Estuary-Delta.

机构信息

Genomic Variation Lab, University of California Davis, One Shields Ave, Davis, CA, 95616, USA.

出版信息

Mol Ecol Resour. 2015 Mar;15(2):278-84. doi: 10.1111/1755-0998.12305. Epub 2014 Aug 13.

Abstract

The effect of predation on native fish by introduced species in the San Francisco Estuary-Delta (SFE) has not been thoroughly studied despite its potential to impact species abundances. Species-specific quantitative PCR (qPCR) is an accurate method for identifying species from exogenous DNA samples. Quantitative PCR assays can be used for detecting prey in gut contents or faeces, discriminating between cryptic species, or detecting rare aquatic species. We designed ten TaqMan qPCR assays for fish species from the SFE watershed most likely to be affected by non-native piscivores. The assays designed are highly specific, producing no signal from co-occurring or related species, and sensitive, with a limit of detection between 3.2 and 0.013 pg/μL of target DNA. These assays will be used in conjunction with a high-throughput qPCR platform to compare predation rates between native and non-native piscivores and assess the impacts of predation in the system.

摘要

尽管捕食对物种数量可能产生影响,但旧金山湾-三角洲(SFE)的本地鱼类受引入物种捕食的影响尚未得到深入研究。物种特异性定量 PCR(qPCR)是一种从外源 DNA 样本中识别物种的准确方法。定量 PCR 分析可用于检测肠道内容物或粪便中的猎物,区分隐存物种,或检测稀有水生物种。我们为最有可能受到非本地食鱼动物影响的 SFE 流域的鱼类设计了十个 TaqMan qPCR 分析。设计的分析具有高度特异性,不会从共存或相关物种产生信号,并且具有高灵敏度,检测限在 3.2 至 0.013 pg/μL 目标 DNA 之间。这些分析将与高通量 qPCR 平台结合使用,以比较本地和非本地食鱼动物的捕食率,并评估该系统中捕食的影响。

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